Guest Column | December 8, 2025

Rats Behind The Wheel: The Right Preclinical Way To Investigate Drug Effects On Driving?

By Anke Rosch, Boehringer Ingelheim Pharma GmbH & Co. KG

Couple driving in car-GettyImages-1320539830

Some time ago, Kelly Lambert1,2 at the University of Richmond, USA, saw her research on how environmental enrichment improves learning of complex behavior like car driving in rats go viral. By remarkable coincidence, a respected colleague asked me what on earth we could do preclinically for drug-induced impairment of driving ability. This column reviews the causes, the regulatory environment, and the limitations of potential preclinical investigations in drug development to study the impact of medications on operating a motor vehicle, accounting for its complex nature.

Background

A Dutch study from 1994,3 showed an about twofold elevated risk of fatal traffic accidents in people taking medical drugs. The synergistic effect of underlying diseases on driving ability complicates its evaluation during clinical drug development.

For example, benzodiazepines4,5,6 and z-drugs (such as zolpidem)7 more than double the risk of car crashes. Their indications, like depression and anxiety8 or insomnia,9 are themselves associated with up to a threefold higher accident probability. Antihistamines are linked to a threefold higher crash risk,10 which is only slightly higher than normal in individuals with allergy symptoms.11 The risk of accidents doubles for chronic pain patients, mirroring12, 13 that of driving on opioids10, 14 and increasing 22-fold when combined with alcohol.15

In the EU, the DRUID study (Driving Under the Influence of Drugs, Alcohol and Medications, 2007-2009)16 provided data on the involvement of alcohol, illicit drugs, and medications in traffic accidents. The results were based on investigations of blood and oral fluid samples supplemented by roadside surveys of 50,000 drivers. The study revealed alcohol as the most prevalent substance (38.9%) in dead traffic victims, followed by benzodiazepines (5.2%), z‑drugs (2.8%), amphetamines (2.1%), and opioids (1.5%), in relation to 34,826 fatal traffic accidents in Europe in 2009.17

Based on an analysis of 7,159 fatal car crashes in the United States Fatality Analysis Reporting System, Brady and Li18 reported drug prevalences for alcohol (35.7%), cannabinoids (9.9% vs. 1.8% in the EU), stimulants (e.g., amphetamine, 9.2%), narcotic analgesics (such as opioids, 5.8%), and depressants (e.g., benzodiazepines, z-drugs, 4%) from 2007 to 2010, representative of 30,296 fatal crashes in total in 2010.19

Conversely, a few studies reported drug-induced improvements in driving ability. Methylphenidate, for example, enhanced driving performance in adolescents and young adults (mean age of 17-18 years) with ADHD,20,21 individuals with Parkinson's disease,22 and those with epilepsy.23

Regulatory Coverage And Demands

The Food and Drug Administration’s Guidance for Evaluating Drug Effects on the Ability to Operate a Motor Vehicle, released in 2017,24 outlines the objectives and principles for investigating driving ability during drug development. Psychoactive but also nonpsychoactive drugs that affect driving indirectly (e.g., antidiabetics and mydriatics) are of major concern. The guidance recommends a tiered approach comprising pharmacological/toxicological, clinical behavioral and epidemiological monitoring (e.g., drug-disease interactions).  The following sections will describe the regulatory requirements and discuss options for a preclinical assessment.

The Neurocognitive Challenge

Driving is a challenging task, dependent on the function of a wide range of cognitive abilities specified in the guidance.24, 25, 26 Publications linked impaired executive function,27, 28, 29 visuospatial skills,28, 29, 30 attention, and processing speed27, 28 to reduced driving ability in older adults (mean of 67-78 years)27 with cognitive deficits, and individuals taking benzodiazepines31 or opioids.32

Functional magnetic resonance imaging (fMRI) showed that driving involves neural brain activity in diverse anatomic structures such as the cerebellum, occipital, parietal, limbic regions, basal ganglia, prefrontal/frontal region, and premotor/motor cortex.33, 34, 35, 36 By managing motor control, action planning, and movement processing, these regions contribute to executive function,37, 38 attention,39, 40, 41 and visuospatial skills.42, 43  Cellularly, the following drug targets play a role: noradrenergic, dopaminergic, serotonergic, acetylcholinergic,14, 44, 45, 46 opioidergic,14 and GABAergic47 neurotransmission, cannabinoid receptors,48 and transporters (SERT, NET and DAT),14 Glycine,49 N-methyl-D-aspartate (NMDA),50 and orexin receptors51. Given the interconnectedness of these structures, definitively linking specific receptors and brain areas to cognitive functions remains impossible, despite extensive research.

An Evaluation of Dedicated Test Scenarios

The FDA Guidance24 requests preclinical investigations into chemical similarity (e.g., to benzodiazepines), on- and off-target receptor binding,52, 53, 54 and pharmacokinetic characteristics such as brain penetration.55 Regulators suggest that the multifaceted nature of driving is only mimicked in in vivo studies. Unfortunately, this level of complexity would consequently call for equally sophisticated study designs when second tier CNS tests are requested.

The set-shifting task in monkeys, rats and mice provides insights into the effects on cognitive flexibility, executive function and attention, involving brain regions such as the prefrontal and frontal cortices and thalamus.56, 57 It resembles human performance in the Wisconsin Card Sorting Test58 and the Cambridge Neuropsychological Test Automated Battery Intra-/Extra-Dimensional Set-Shift Task (ID/ED).59 During the set-shifting task (an operant procedure), animals must adjust their learned responses to novel stimuli combinations within one dimension such as changing from house light to cue lights above the levers60 or across different dimensions like from color to shape61 to earn a food reward. Measured parameters comprise the number of errors committed and trials required to achieve the predefined criterion for set-shifting/response discrimination and reversal learning.

Carli et al.62 developed the Five-Choice Serial Reaction Time (5-CSRT) task in rats, modified for mice,63 monkeys,64 and zebrafish.65 This paradigm is based on the continuous performance tests in humans.66 The original apparatus for rats consists of a chamber with a curved front wall containing five active horizontally placed holes. On the opposite wall, a food receptacle is connected to an automatic dispenser. An infrared light beam, whose fixed-frequency light stimulus can be modified according to time length, is associated with each hole. After a training phase of several weeks, the animal is placed in front of the curved wall. After each visual (light) stimulus, the animal has a limited time to respond with a nose poke in the illuminated hole out of the five locations. A correct response results in an immediate reward with a food pellet at the opposite wall. Parameters such as the accuracy rate of responding, premature responding, and response latencies allow conclusions on sustained attention, processing speed, executive function, and impulse control associated with the integrity of frontal, parietal, prefrontal and cingulate cortices, and the limbic region.67, 68

The Cincinnati Water Maze for rodents, adapted from the Biel Water Maze69 by Vorhees,70 is a more feasible operant procedure used for learning and memory investigations in rats and mice. The partially submerged multiple T-maze utilized for the task contains dead-end and open pathways and facilitates testing under light and dark conditions. During testing, the animal has to navigate from the starting point to the hidden escape platform by using internal (in the light and dark) and external (during light) cues after a training period of five under light conditions or 15 days in the dark. Measures of escape latency and number of path errors indicate impacts on spatial learning, visuospatial skills, memory and key brain structures such as the basal ganglia, prefrontal cortex, thalamus, and limbic region.71

Conclusions And Perspectives

Preclinical investigation of drug-induced effects on driving ability is sufficiently addressed ex vivo by chemical similarity, secondary pharmacology, and pharmacokinetic studies in case an abuse potential assessment must be conducted in parallel.72, 73

Regulatory in vivo tests such as the modified Irwin test74 or the functional observation battery in rats or mice,75 performed as a stand-alone safety pharmacology study76 or implemented in toxicity studies, fail to investigate learning and memory.77 In addition, they show low predictivity for clinical adverse events.78

The set-shifting task and other intricate behavioral models are non-standardizable research tools that are valuable for exploring the dynamic interplay of perception, decision-making, and motor control but are of uncertain value for assessing driving ability. The Cincinnati Water Maze for rats, despite its methodological complexity, has gained regulators‘ acceptance for investigation of neurodevelopmental toxicity in extended one-generation reproductive toxicity studies (EOGRTS) of chemicals, as reflected in the Organization for Economic Co-operation and Development (OECD) test guidelines 443 and 426.79, 80 In case of a concern about thyroid toxicity,81 the European Chemical Agency (ECHA) started to request tests on learning and memory in 2022.82 Although the Cincinnati Water Maze could potentially be integrated into rodent toxicity studies, the time-consuming animal training and the need to coordinate the tests with regular study activities will prevent it from becoming standard.

Alternative methods, such as fMRI83 cannot be routinely integrated in regulatory studies; they pose a cost burden and require a high level of continuous qualification and validation. Biomarkers are currently not validated in diseased humans‑not to mention in healthy animals. They are more closely associated with the pathology of the diseases than with functional aspects of learning and memory. Biomarkers involved in neurodegeneration, cognitive decline, memory and plaque formation such as amyloid-β 1 to 42 peptide and the microtubule-associated protein t-tau,84, 85 the neuron-specific cytoskeletal protein neurofilament light chain (NfL),86, 87, 88 and the brain-derived neurotrophic factor89, 90, 91 deserve special mention here.

Taken all together, our current mandatory preclinical drug development activities provide only limited but often non-specific signals that may indicate drug-induced effects on driving ability. Second-tier in vivo tests addressing more complex aspects of driving stand in contrast to the progressive phase-out of animal studies. Metaphorically, even a car-driving rat could not reliably tell us if people on medications can operate motor vehicles safely as such behavior is not part of its natural repertoire. While further refinement of safety and toxicological studies may add value, the most decisive step in evaluating drug-induced driving impairment remains, in my view, clinical trials.

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About The Author

Anke Rosch is a board-certified pharmacologist and toxicologist working at Boehringer Ingelheim Pharma GmbH & Co. KG. A doctor of veterinary medicine, she has more than 20 years of experience in the pharmaceutical industry and has special expertise in safety pharmacology. Anke can be reached at ankerosch.PharmacolTox@t-online.de.